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  • 1985-1989  (585)
  • 1975-1979
  • 1840-1849
  • 1986  (585)
  • Biochemistry and Biotechnology  (519)
  • Ultrastructure
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    European archives of oto-rhino-laryngology and head & neck 243 (1986), S. 121-125 
    ISSN: 1434-4726
    Keywords: Bacterial labyrinthitis ; Ultrastructure ; Mycobacterium ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We used electron microscopy to investigate Mycobacterium fortuitum -induced changes in the inner ears of mice. We found that the inner and outer hair cells had degenerated and disappeared in the organ of Corti. Changes in the lower turn of the cochlea were more severe than those of the upper turn while the changes of the outer hair cells were more severe than those of the inner hair cells. Disappearance, fusion and ballooning of the sensory hairs were observed in the vestibular organs. The bacterial extract also induced inner ear damage which was similar to that caused by live M. fortuitum.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    European archives of oto-rhino-laryngology and head & neck 243 (1986), S. 260-264 
    ISSN: 1434-4726
    Keywords: Argyria ; Chemosensory function ; Oral Mucosa ; Ultrastructure ; X-ray microanalysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Generalized argyrosis can produce a number of abnormalities, including skin discoloration, liver and kidney dysfunction. We describe a patient with generalized argyrosis following long-term self-treatment with oral silver intake, in whom skin discoloration, progressive taste and smell disorders, vertigo and hypesthesia were observed. These findings were confirmed by chemosensory tests and electrophysiological investigations. The development of hypogeusia was assessed by subjective tests, while the progression of hyposmia was followed by recording olfactory evoked cortical potentials. Light and electron microscopy of tissue samplings demonstrated electron-dense mineral deposits in basal membranes, in macrophages, in the perineurium of peripheral nerves, along elastic and collagenous fibers, and in necrotic cells of the oral submucosa. Silver and sulfur deposits in affected tissues could be defined by X-ray microanalysis. The quantitative ratio between silver and sulfur in involved tissues was similar to that of an inorganic silver-sulfide (Ag2S) standard. The minute increase in the sulfur content when compared to the inorganic standard suggested a sulfur containing organic matrix of the tissue precipitates. Our findings indicate that the affinity of silver for membrane and neuronal structures and the deposition of silver as an insoluble compound (Ag2S) induce the progression of clinical disease.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0584
    Keywords: Karyotype ; Bladder carcinoma ; Ultrastructure ; CSF ; Cell line
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The cell line 5637 which originated from a human urinary bladder carcinoma is known to produce GM-CSF and Multi-CSF ectopically. Determination of cell surface antigens defined by monoclonal antibodies was recently reported [6]. Here we report on the ultrastructure and karyology of this CSF secreting cell line. At the ultrastructural level the monolayer in vitro culture and the solid tumors formed in nude mice showed all characteristics consistent with a well-differentiated transitional cell carcinoma (TCC). A subclone was found to grow in suspension and did not secrete any CSF activity. High resolution chromsome analysis revealed chromosomal abnormalities which agreed only in few particulars with nonrandom chromosomal aberrations usually found in TCC. Analysis of the cytogenetic results showed that nearly all structural abnormalities present are known to be associated with acute or chronic human leukemia. The possibility that the ectopic production of CSF in this cell line may be correlated to one or more of the described chromosomal aberrations is discussed.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Archives of dermatological research 278 (1986), S. 398-406 
    ISSN: 1432-069X
    Keywords: Nevocytic nevi ; Congenital ; Ultrastructure ; Nevus cells ; Nerve contacts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary As only few previous investigations considered size-dependent differences in the histology of congenital nevocytic nevi (CNN) or focused on their ultrastructure, a light- and electron-microscopic study of six giant and six medium-sized CNN was performed. Histologically, giant CNN showed minimal junctional nevus elements and a pandermal, diffuse nevus cell extension including the subcutaneous tissue, while medium-sized CNN exhibited more abundant junctional nests and a rather focal pattern of nevus cell extension into the reticular dermis. Ultrastructurally, giant CNN often showed nevus cells with irregular and indented nuclei and also nevus cells with highly complicated dendrites. Nuclear inclusions, multiple cilia and centrioles, and collagen phagocytosis in nevus cells were found in both groups of CNN. Direct contacts between nevus cells and nerve fibers were observed in mediumsized CNN only. The findings provided no clear morphological indication why giant CNN have a higher risk for malignant degeneration than medium-sized CNN or acquired nevocytic nevi (ANN). On the other hand, the histological differences speak in favor of a different ontogenesis of the various size classes of CNN and ANN.
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  • 5
    ISSN: 1432-2307
    Keywords: Epithelioid haemangioendothelioma ; IVBAT ; Immunohistochemistry ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Three patients with epithelioid haemangioendothelioma (EHE) are described. Two patients presented with pulmonary infiltrates and one with a hepatic tumour. All had a metastatic disease ending fatally, and all were autopsied. The diagnosis was confirmed either by immunohistological or ultrastructural analysis. All three tumours were cytokeratin-negative and vimentin-positive, while only two contained cells reacting with the antibody of factor VIII-related antigen. Electron microscopy of the third tumour revealed features indicating endothelial differentation. A short literature review is also presented demonstrating that the outlook of EHE is worse than previously thought.
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  • 6
    ISSN: 1432-2307
    Keywords: Mesothelioma ; Serous membrane ; Localized tumour ; Histogenesis ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary It is uncertain whether localized lesions of serosal membranes have a kinship to mesotheliomas or are truly fibromatous in nature. Ultrastructural and immunohistochemical investigations were carried out on 12 localized benign and malignant pleural and peritoneal tumours from 10 patients. Electron microscopic findings, including the consistent and non-fibroblastic cellular organization of localized neoplasms, the presence of some form of intercellular junctions in 7 of 10 cases, basal lamina deposition in 3 cases, and polarized microvilli in one case indicated a form of mesothelial differentiation. Using monoclonal and polyclonal antibodies, positive immunostaining of tumour cells for cytokeratin peptides was detected in one case, while antibody to vimentin stained four cases. Light microscopic, ultrastructural and immunohistochemical features of one benign localized serosal tumour, with a unique blend of epithelial and spindle cells, provided further evidence for a histogenic link between localized serosal tumours and diffuse epithelial mesotheliomas. On the basis of the current findings and reports in the literature, it would appear that the majority of localized tumours of serosal membranes are a subset of mesothelioma, while a minority are fibromas.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Virchows Archiv 410 (1986), S. 43-48 
    ISSN: 1432-2307
    Keywords: Endocarditis ; Staphylococcus ; Colonisation ; Morphology ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The initial colonization, byStaphylococcus aureus, of the catheter damaged aortic valve and aorta of the rabbit, was examined by light and electron microscopy at 15 min, 3 h and 24 h post inoculation (PI). At 15 min PI, the majority of bacteria (80%) were located on the lateral surfaces of the thrombic vegetations while 20% were attached directly to the connective tissue of the aortic valve and aorta in areas where the endothelial lining was disrupted. By 3 h the bacteria on the thrombic vegetations were covered by fibrin. At this time, the bacteria both within the vegetations and on the surface of the vasculature were undergoing multiplication to form small groups. The precipitation of thrombus around the bacteria attached to the surface of the aorta to form microscopic infected vegetations had occurred by 24 h PI. The colonizing bacteria did not elicit any phagocytic response. The colonization of the cardiovasculature byStaph. aureus did not necessarily require pre-existing vegetations.
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  • 8
    ISSN: 1432-2307
    Keywords: Human acute pancreatitis ; Enzyme immunocytochemistry ; Ultrastructure ; Acinar cell degranulation ; Fat necrosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Human acute pancreatitis results from an autodigestive process frequently associated with alcohol abuse, gall stone disease and shock. Peripancreatic fat necrosis was identified as one of the earliest visible lesions, whereas acinar cell necrosis and haemorrhage were regarded as secondary changes. To examine the alterations in acinar cells in more detail, their enzyme content and fine structural features were studied immunocytochemically using antisera against α-amylase, lipase, trypsin, chymotrypsin and pancreatic stone protein, and electronmicroscopically in pancreatic tissues from patients with severe acute pancreatitis. Peripheral acinar cells in the immediate vicinity of fat necrosis were found to be heavily degranulated, while acinar cells at some distance of necrosis fully retained their enzyme content. Other frequent changes of the acinar cells included cuboidal transformation, loss of microvilli, increased occurrence of autophagosomes, and formation of enlarged acinar lumina. As there was no apparent cell membrane leakage or rupture of duct lumina, it is concluded that the acinar cells adjacent to fat necrosis release their granules by undirected basolateral extrusion. The findings thus suggest that one of the basic defects in acute pancreatitis is the uncontrolled release of enzymes from peripheral acinar cells into the interstitial space which, in turn, presumably by the action of lipase, leads to autodigestive fat necrosis.
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  • 9
    ISSN: 1434-0879
    Keywords: Monkey spermatozoa ; Gossypol effect ; In vivo examination ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The present study examines the ultrastructure of ejaculated spermatoza from bonnet monkey, Macaca radiata under noraml conditions, with gossypol treatment and during recovery from such treatment. Monkeys were fed orally with gossypol acetic acid (GAA) for 3 months (4 mg/monkey/5 days a weak). Semen samples collected by electroejaculation, and the spermatozoa were examined using both light and electron microscopy. The degree of motility was also noted by Kalla et al. [12]. Ejaculated spermatoza were immotile 90 days after GAA treatment, but little evidence for any abnormality in the spermatozoa could be seen by light microscopy. Some ultrastructural changes were observed, but not to the extent previously reported in spermatozoa of Macaca fascicularis [23]. After termination of treatment, semen samples were obtained every 5th day until sperm count and motility recovered to the normal level. After 90 days only a small proportion of spermatozoa showed abnormal structure. We conclude that in a subhuman animal model gossypol induced effects on sperm motility and morphology are reversible.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 173 (1986), S. 295-298 
    ISSN: 1432-0568
    Keywords: Cardiodilatin ; Peptide Hormone ; Heart ; Human ; Immunohistochemistry ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The immunogold technique was applied to detect cardiodilatin-immunoreactivity (CDD-IR) in human heart. Using antibodies directed against C-terminal regions of CDD, we could identify colloidal gold label in specific atrial granules of human heart. The granules in all areas of the myoendocrine cells were homogeneously labelled and all right atrial biopsy specimen of patients with different heart diseases reacted positively and had the same distribution of label. The Golgi-apparatus and ergastoplasma exhibited and extremely weak CDD-IR. The results indicate a predominant storage of CDD in specific granules.
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  • 11
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 173 (1986), S. 361-370 
    ISSN: 1432-0568
    Keywords: Electrical synapses ; Chemical synapses ; Ultrastructure ; Morphometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Thin sectioning and freeze-fracturing have revealed the distribution of gap junctions and chemical synapses in the synaptic interface of the large myelinated club endings on the lateral dendrite of the goldfish Mauthner cell. In 12 samples of club endings fractured completely or nearly completely, the apposed synaptic membrane area averaged 39.090 μm2, of which 16.6% was occupied by gap junctions and about 4 to 5% by the active zones of chemical synapses. The numerical profile density (number per unit area of the synaptic membrane) of gap junctions varied greatly, from 1.78 to 6.30, and was mostly in inverse proportion to their size. The chemical synapses were located mainly in two places: in the circumferential rim of the synaptic membrane next to the widened extracellular space, and in the margins of intraterminal invaginations of the synaptic cleft. The axoplasm of the preterminal axon, just after losing its myelin sheath, was filled with microtubules, among which neurofilaments gathered into many small bundles. The correlation between the areas of gap junctions and the chemical synapses and the amplitude of the excitatory postsynaptic potentials (EPSP) is discussed.
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  • 12
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 173 (1986), S. 385-391 
    ISSN: 1432-0568
    Keywords: Elastic cartilage ; Chondrogenesis ; External ear ; Rat ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Selected ultrastructural features of chondrocytes and the extracellular matrix in the developing elastic cartilage of the external ear were studied in rat fetuses and young animals. The cytoplasmic lipid droplets were first observed in the 19-day fetus. They increase in number and size during the first post-natal week. The elastogenesis proceeds in the sequence: oxytalan fibers (17-day fetus), elaunin fibers (1-day rat), elastic fibers (5-day rat). Intermediary stages between the randomly oriented individual microfibrils and bundles of microfibrils (oxytalan fibers) were also observed.
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  • 13
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 174 (1986), S. 27-33 
    ISSN: 1432-0568
    Keywords: Cerebral cortex ; Antisera ; Biopsy ; Postmortem Alzheimer's type dementia ; Ultrastructure ; Epilepsy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary NPY-containing neuronal structures in the cereoral cortex of surgical tissue samples were compared to those in postmortem material by immunocytochemical methods. However, the quality of preservation of individual neurons and axonal and dendritic plexuses in the neuropil is unusually fine in the surgical specimens. This result is most likely attributable to the excellent fixation that can be regularly achieved by rapid and careful handling of tissue during and after surgical removal. The tissue is suitable for both light and electron microscopy, and the superior preservation also leads to intense, reliable antibody reactions. Postmortem tissue samples can provide good specimens for immunocytochemistry when properly handled as previously described. However the postmortem delays prior to fixation disrupt neuronal integrity in the immunostained structures. Nevertheless, postmortem material from carefully studied subjects of neurological diseases compared with age matched controls can provide valuable information.
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  • 14
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 173 (1986), S. 317-322 
    ISSN: 1432-0568
    Keywords: Paneth cell ; Ultrastructure ; Morphometry ; Germfree rats ; Ex-germfree rats
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Ultrastructural changes of Paneth cells of germfree (Gf) rats which had been inoculated with bacteria-containing feces from conventionally-reared (SPF) rats were quantitatively examined. 12 and 24 h after inoculation, the Paneth cells showed a striking decrease in the number of secretory granules and the occurrence of large vacuoles. Phagosomes containing bacteria were not seen. After 4 days, the secretory granules reaccumulated and smooth-surfaced apical vesicles increased in number. It is discussed that the large vacuoles may be related to membrane-retrieval events following the massive extrusion of secretory granules whereas the apical vesicles appear to serve this function when exocytosis is not pronounced. In addition to the large secretory granules ca. 10% of Paneth cell profiles contained a few dense-cored vesicles measuring about 150 nm in diameter which resemble peptidergic neurosecretory granules.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Acta neuropathologica 71 (1986), S. 83-87 
    ISSN: 1432-0533
    Keywords: Astrocytes ; Brain neoplasms ; Oligodendroglioma ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Ultrastructural concentric laminations have previously been thought to be specific to oligodendroglioma. However, these structures were also recognized in fibrillary astrocytomas, a mixed glioma and a glioblastoma. These laminations continued or closely related to attenuated processes or cytoplasm of astrocytic tumor cells. In addition, some lamellae contained glial filaments. It is considered that the concentric laminations are derived from attenuated astrocytic processes and have no relationship with myelin.
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  • 16
    Electronic Resource
    Electronic Resource
    Springer
    Acta neuropathologica 70 (1986), S. 44-52 
    ISSN: 1432-0533
    Keywords: Gliosarcoma ; Myxoid tumor ; Extracranial metastasis ; Ultrastructure ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The case of a 22-year-old female suffering from glioblastoma with sarcomatous component (WHO; gliosarcoma) is presented. The tumor consisted of glioblastomatous and sarcomatouos components, and in part of the pleomorphic astrocytomatous region of the glioblastoma there was a prominent production of myxoid matrix. On the basis of its histochemical and immunohistochemical characteristics, the present case strongly suggested that glial cells of neuroectodermal origin assume part of the responsibility for production of myxoid material.
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  • 17
    Electronic Resource
    Electronic Resource
    Springer
    Acta neuropathologica 72 (1986), S. 134-141 
    ISSN: 1432-0533
    Keywords: Cerebral embolism ; Fungal vasculitis ; Paecilomyces javanicus ; Prosthetic valve endocarditis ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A 41-year-old diabetic woman developed Paecilomyces javanicus endocarditis and subsequent cerebral fungal embolism and vasculitis with massive infarction and hemorrhage 6 years after a mitral valve replacement with porcine heterograft. The organism was identified by the culture from the infected brain tissue and aortic valve. Cerebral infection due to fungus Paecilomyces is rare. This report documents the cerebral manifestations of fungus Paecilomyces javanicus and describes for the first time the ultrastructure of such an organism obtained from the infected tissue. Their ultrastructural findings, similar to those described in Paecilomyces farinosus derived from culture, include a double-layered cell wall, a triple unit plasma membrane, abundant cytoplasmic lipid droplets, vacuoles, membranous profiles and septal formation. This report further demonstrates hyphae with packed 50A filaments and granulofibrillary material and features suggestive of intrahyphal hyphae, not previously described in fungus Paecilomyces.
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  • 18
    Electronic Resource
    Electronic Resource
    Springer
    Acta neuropathologica 70 (1986), S. 127-134 
    ISSN: 1432-0533
    Keywords: Avian muscle spindle ; Denervation ; Intrafusal fiber ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The role of the nerve in maintaining the ultrastructural integrity of avian muscle spindles was investigated by denervating the pigcon's extensor digitorum communis for periods of 10, 19, and 28 days. The equatorial region of control intrafusal fibers had a reduced density of myofilaments. Sensory endings contained mitochondria and structures resembling synaptic vesicles, and were associated with satellite cells. In the polar region, fibers had a high concentration of myofilaments; small motor endings, unlike sensory endings, lay outside of the fiber's basal lamina. The outer capsule consisted of thin, tightly layered cells which gradually became reduced in number distal to the equatorial region. In both equatorial and polar regions the capsule became more disrupted with longer denervation periods, and lysosomes and phagocytes became more abundant. The equatorial region of denervated fibers contained many myofibrils and some had peripherally-located nuclei, unlike the controls; sensory terminals were absent. The polar region of some fibers had disorganized myofilaments and others had a reduced myofilament density. Fiber diameters increased significantly in both regions. Thus, denervated intrafusal fibers lost some characteristics which distinguish them from extrafusal fibers.
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  • 19
    Electronic Resource
    Electronic Resource
    Springer
    Acta neuropathologica 70 (1986), S. 71-74 
    ISSN: 1432-0533
    Keywords: 6-Aminonicotinamide ; Ependymal cell ; Mitosis ; Suckling mice ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Mitotic ependymal cells were encountered in 10-day-old mice treated with 6-aminonicotinamide, an antagonist of niacin. These occurred along the medial surface of the lateral ventricle and the ventral portion of the aqueduct. Electron microscopy revealed that both mitotic ependymal cells had eccentrically placed chromosomes without a nuclear membrane and well-formed gap junctions in contact with adjacent ependymal cells. Microtubules from a centriole radiated to the chromosomes. These data show that cell division occurs in morphologically matured ependymal cells in the postnatal brain under pathological conditions. We believe this to be the first ultrastructural demonstration of this phenomenon.
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  • 20
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 174 (1986), S. 379-389 
    ISSN: 1432-0568
    Keywords: Lungs ; Amphibia ; Ultrastructure ; Morphometry ; Surfactant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Lungs ofHyla arborea L. were examined by scanning and transmission electron microscopy and morphometric methods. The lungs contain several interconnected folds in a netlike reticular arrangement of first, second and third order, mainly covered with pneumocytes. On the septa of first and second order, irregularly distributed small patches of ciliated epithelium devoid of goblet cells are located. Dome-shaped neuroepithelial bodies can be seen in the vicinity of ciliated epithelium. The pulmonary epithelium consists of one type of pneumocyte, which contain in their cytoplasm three kinds of bodies: lamellar, dense and multivesicular. The dense bodies are precursors of lamellar bodies (LBs), while the multivesicular bodies are incorporated into the LBs, being later secreted to the air space. The lining layer covering the internal lung surface contains numerous transformed LBs but tubular myelin figures are scarce. The surface of the lining layer is coated by a thin film. The air-blood barrier, consisting of three layers: epithelium, interstitial space and endothelium, is 0.6 μm thick.
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  • 21
    Electronic Resource
    Electronic Resource
    Springer
    Acta neuropathologica 69 (1986), S. 81-90 
    ISSN: 1432-0533
    Keywords: Creutzfeldt-Jacob disease ; Slow virus infection ; Subacute spongiform encephalopathy ; Ultrastructure ; Synapse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Guinea pigs inoculated with brain homogenate from serially passaged Creutzfeldt-Jakob disease (CJD) were killed biweekly starting at week 2 until terminal illness (about 200 days following inoculation). A mild swelling of postsynaptic dendrites and an increase in the number of glial filaments in astrocytic processes was seen at week 4, followed by increased swelling and lucency of axons and dendrites by week 6 post inoculation (p.i.). Severe undulation and focal interruptions of synaptic membranes were also observed both at weeks 4 and 6. By week 8, one could see cystically dilated cellular processes. These sometimes showed continuity with adjacent swollen processes through focally disrupted plasma membranes, and most likely represent a progressive enlargement of vacuoles through fusion and subsequent addition of adjoining processes. The spongiform changes increased mildly between week 8 and week 10 and remained essentially the same in subsequent weeks. After week 24 there was a sharp increase in both the number and size of vacuoles. At week 24 severe structural alterations were present both in the neurons and astrocytes, and numerous intranuclear inclusions were demonstrated in many neuronal nuclei. This study shows that morphological changes in the brain occur considerably earlier than the clinical manifestations of the disease. In the early phase of the disease, there were significant alterations on the dendrites and synapses.
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  • 22
    ISSN: 1432-0533
    Keywords: Streptozotocin-diabetic rat ; Myenteric plexus ; VIP-ergic nerves ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Enteric nerves in the ileum of rats 8 weeks after streptozotocin-induction of diabetes were examined under the electron microscope before and after immunolabeling for vasoactive intestinal polypeptide (VIP). These studies have provided evidence of degenerative changes in the myenteric nerve fibres of diabetic rats, many of which were shown to contain VIP. It is suggested that VIP-ergic nerves in the gut may play a role in the developent of gastrointestinal dysfunction in diabetes.
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  • 23
    ISSN: 1432-0533
    Keywords: Brain tumor ; Rhabdomyosarcoma ; Immunohistochemistry ; Ultrastructure ; Cerebral paragonimiasis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A necropsy case of a primary rhabdomyosarcoma with chronic paragonimiasis in the cerebrum of a 68-year-old man is reported. The clinical data showed a right hemiplegia and dysarthria which became lethal in 6 months even though operation and radiation therapy were performed. Computed tomography revealed a large low-density area associated with the peripheral enhancement in the left basal ganglia, and multiple conglomerated calcified masses in the left temporal and occipital lobes. Biopsied and necropsied materials of the tumor in the basal ganglia was reddish brown in color and histologically was composed of purely mesenchymal derivatives with both embryonal and mature striated muscle cells but neither neuronal nor glial elements. Some of the tumor cells with extending slender cytoplasms showed obvious cross striations at the light and electron microscope levels and immunohistochemical reactivity for myoglobin. All tumor cells were also positive for vimentin, but not for glial fibrillary acidic protein. The clinical and necropsy findings revealed no primary lesion anywhere but in the brain. In addition, numerous dead oval eggs ofParagonimus westermani were found in many cystoid lesions encapsulated by thick connective tissues with calcification and/or ossification. Clinicopathological features of 24 cases of primary rhabdomyosarcoma of the central nervous system reported in the literature are reviewed briefly. The histogenesis of this tumor are discussed together with comments on cerebral paragonimiasis.
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  • 24
    Electronic Resource
    Electronic Resource
    Springer
    Archives of dermatological research 279 (1986), S. 77-82 
    ISSN: 1432-069X
    Keywords: Lichen nitidus ; Lichen plahus ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Lichen nitidus (LN) and lichen planus (LP) are considered by some investigators to be two variants of the same disease, and by others to be two distinct dermatoses. In order to obtain further information about the relationship between LN and LP we examined the ultrastructure of lesions from two LN patients. In the central part of the lesion, the basement membrane was absent, or was interrupted by migrating phagocytes or lymphocytes. The basal cells and the lower cells of the stratum spinosum exhibited karyolysis and appeared to be compressed and often necrotic. In the upper dermis irregular cell debris full of clumps of tonofilaments and colloid-body-like structures was observed. A dense dermal infiltrate of macrophages, lymphocytes, fibrocytes, and Sezary-like cells was present. Signs of cooperation between lymphocytes and macrophages were also evident. The periphery of the lesion showed no pathological features except for enlargement of the intercellular spaces and the presence of mononuclear cells scattered through the epidermis. Several normal Langerhans cells were noticed. These ultrastructural findings were quite similar to those reported for LP.
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  • 25
    Electronic Resource
    Electronic Resource
    Springer
    Cellular and molecular life sciences 42 (1986), S. 144-147 
    ISSN: 1420-9071
    Keywords: Ultrastructure ; catalase ; D-amino acid oxidase ; fetal mouse liver ; hepatocytes ; peroxisomes ; muscular dysgenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the hepatocytes of ‘normal’ fetal mice from mothers which were carriers of muscular dysgenesis, catalase and D-amino acid oxidase (DAAO) positive as well as negative peroxisomes were observed. DAAO reaction product was occasionally localized in patches around cell membranes and DAAO-positive peroxisomes were frequently observed near mitochondria.
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  • 26
    ISSN: 1432-1106
    Keywords: Optic tectum ; Neurotransmitters ; Cholinergic markers ; Ultrastructure ; Goldfish
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Neurochemical parameters associated with cholinergic and excitatory amino acid transmission, were measured in synaptosomes of the goldfish optic tectum at different times after unilateral eye ablation. Significant decreases in choline acetyltransferase and acetylcholinesterase were measured 12 and 30 days after enucleation. The high affinity choline uptake did not parallel the decrease in cholinergic enzymes. Instead there was a significant increase of the uptake per unit of protein (though not relative to the total number of tectal synaptosomes). No decrease of the high affinity D-3H aspartate uptake was measured in the deafferentated optic tectum. Electron microscopic observations showed a correspondence between the time course of cholinergic enzyme decrease and the degeneration of retinal afferents to the tectum. The present results support the notion that acetylcholine is a better candidate than the excitatory amino acids for a neurotransmitter role in the fish optic tectum.
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  • 27
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    European journal of pediatrics 144 (1986), S. 557-562 
    ISSN: 1432-1076
    Keywords: Rimmed vacuoles ; Childhood ; Myopathy ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A 5-year-old boy suffered from a slowly progressive non-familial neuromuscular disease, clinically marked by generalised muscle weakness, atrophy and hypotonia, a “myopathic” EMG and mildly elevated CK values. His gastrocnemius muscle showed marked myopathy, type I fibre predominance, and numerous “rimmed” vacuoles. This boy's condition is regarded as a childhood neuromuscular disease with rimmed vacuoles.
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  • 28
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    Child's nervous system 2 (1986), S. 297-300 
    ISSN: 1433-0350
    Keywords: Pineocytoma ; Magnetic resonance imaging ; Computed tomography ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The radiological and pathological findings in three children with pineocytomas are described. The patients presented with symptoms and signs of increased intracranial pressure. CT scans demonstrated obstructive hydrocephalus, but failed to visualize pineal masses that were subsequently seen on MR scans. A suboccipital, supracerebellar approach was used to obtain tissue for histological diagnosis and, in one case, achieve a total removal. Light microscopy demonstrated typical pineocytomatous rosettes without evidence of gangliocytic or astrocytic differentiation. Electron microscopy revealed numerous dense core vesicles with synaptic ribbons, suggesting neurosecretory capability. These three cases demonstrate the diagnostic advantage of MR imaging for tumors in the pineal region as well as the relatively well-differentiated histopathological characteristics of pineocytomas.
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  • 29
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    Virchows Archiv 409 (1986), S. 163-169 
    ISSN: 1432-2307
    Keywords: Thymoma ; Thymic carcinoma ; Ultrastructure ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A case of thymic carcinoma arising within a lymphocyte rich thymoma is reported. The undifferentiated carcinoma contained cellular elements resembling choriocarcinoma but could be differentiated there-from by positive staining for prekeratin antigen and an absence of staining for B-HCG antigen utilizing immunohistochemical techniques.
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  • 30
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    Theoretical and applied genetics 72 (1986), S. 840-844 
    ISSN: 1432-2242
    Keywords: Chlorophytum comosum ; First pollen mitosis ; Male plastid inheritance ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The behaviour of plastids and mitochondria during the formation and development of the male gametophyte of Chlorophytum comosum has been investigated using electron microscopy. During first pollen mitosis an intracellular polarization of plastids occurs in that the plastids are clustered in the centre of the microspore. The originating generative cell normally lacks plastids. Only in a small number of microspores have plastids been observed near the dividing nucleus of the microspore and later on in the generative cell. These observations agree with the genetic investigations of Collins (1922) on the mode of plastid inheritance which demonstrated a small amount of biparental plastid inheritance in Chlorophytum. The cytological mechanisms underlying plastid polarization during the first pollen mitosis are discussed.
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  • 31
    ISSN: 1432-0878
    Keywords: Glucocorticoids ; Cartilage ; Growth ; Histochemistry ; Ultrastructure ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effects of different doses of various steroids on growth, and on costal and epiphyseal chondrocytes, have been studied in prenatal, immature, and adult Long-Evans rats using histochemical techniques, and both light and electron microscopy. Both prenatal and postnatal treatments have been employed. The steroids used were cortisone (CA), betamethasome (BM), and, in the prenatal group only, dexamethasone (DM). Body weight is reduced in all treated rats (except the low dose of CA) by day 17 of gestation, with greater weight reductions occurring in rats receiving the higher dose level of each steroid. In rats treated prenatally or neonatally, and sacrificed postnatally on days 39–43 or days 116–127, body weights, and tibial and tail lengths, are less than in correspondingly aged controls, thus showing a persistence of the effects of treatment. Costal and epiphyseal cartilages in prenatal rats show cellular, synthetic, and ultrastructural alterations induced by treatment with glucocorticoids but the responses are not necessarily comparable. Except for the low dose of DM, the higher doses of each steroid are more effective in inhibiting, or altering, growth and cellular differentiation in the developing fetuses. Surprisingly, a low dose of DM has a more devastating effect on the cells and extracellular matrix of both costal and epiphyseal cartilage, than do higher dose-levels of the various steroids. Low doses of CA and BM are also effective in inhibiting or altering growth and cellular differentiation, but their effectiveness is largely limited to 17 days of gestation. The order of effect of the various doses of the different steroids on fetal cartilage, listed in decreasing order of severity, is as follows: 0.12 DM, 0.24 DM, 0.42 BM, 50 CA, with 25 CA and 0.18 BM being approximately equal and only slightly different from control cartilages. The effect of prenatal or neonatal glucocorticoid treatment on chondrocytes is minimal in the 30–43 day, or 116–127 day, postnatal groups. In immature and adult rats, cortisone affects the chondrocytes more deleteriously than does betamethasone, and a 5.0 mg dose of CA seems to affect chondrocytes, body weight, and tibial and tail lengths more than 0.2 or 7.5 mg doses.
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  • 32
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    Cell & tissue research 244 (1986), S. 457-466 
    ISSN: 1432-0878
    Keywords: Microthrix ; Microvillus ; Ultrastructure ; Fixation ; Tapeworm ; Hymenolepis diminuta
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of microtriches of the rat tapeworm, Hymenolepis diminuta, was examined with a number of electron-microscopic techniques. Fixatives containing different buffers, non-ionic detergents, chelators, tannic acid and various concentrations of aldehydes were tested for ability to stabilize cytoskeletal components while extracting background material. These methods revealed features unique to these specialized microvilli, and permitted construction of a detailed model of microthrix architecture. The microtriches of H. diminuta are comprised of a microfilament-containing base, a dense cap and a complex junctional region between the base and cap. The microfilaments of the base are contiguous distally with a tubular structure (the junctional tubule) within the junctional region; proximally, the microfilaments end abruptly: a terminal web appears to be absent. A beveled bilayered cylinder of dense material (the core tunic) encircles the microfilamentous core. The core tunics and junctional tubules of the microtriches are specifically and uniformly aligned along the strobila. Microtriches therefore can be distinguished from other microvilli (e.g., those of enterocyte brush borders) by their complex ultrastructure and precise orientation upon the cytoplasmic surface.
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  • 33
    ISSN: 1432-0878
    Keywords: Duodenum ; Cholecystokinin ; Gastrin ; Immunocytochemistry ; Ultrastructure ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Human duodenal endocrine cells reactive with antibodies to cholecystokinin (CCK) 33 (10–20) and/or gastrin 34 (1–15) were studied by a combination of immunohistochemical and electron-microscopic methods. By immunohistochemistry, three types of endocrine cells were distinguished in human duodenal mucosa, i.e., those only positive for only CCK, those positive for both CCK and gastrin and those only positive for only gastrin. Ultrastructurally, the first cell type is characterized by many secretory granules with an eccentric dense core (mean diameter; 271+-74 nm). The second cell type, which was less frequent than the other two, has ultrastructural features that resemble type-I cells. The last cell type was composed of two types of cells containing small secretory granules identical to those of IG cells (mean diameter; 171+-31 nm) or large secretory granules indistinguishable from those of I cells (mean diameter; 286+-50 nm).
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  • 34
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    Cell & tissue research 246 (1986), S. 39-44 
    ISSN: 1432-0878
    Keywords: Ultimobranchial glands ; Cysts ; C-cells ; Ultrastructure ; Chicken (Gallus gallus)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultimobranchial glands of the chicken were examined by electron microscopy and immunocytochemistry using a calcitonin antiserum. Electron microscopy confirmed the presence of C-cells, containing numerous secretory granules storing calcitonin, in the luminal lining of cyst-like structures found in these glands. These cells were furnished with prominent microvillar projections at their luminal surface, and the cytoplasm of the apical region was filled with fibril material. Furthermore, the cells contained prominent junctional complexes and desmosomes at their apico-lateral surfaces. In these C-cells, secretory granules were concentrated near the lumen and some were attached to the apical cell membrane. The luminal content of the cysts had a colloid-like and flocculent appearance, and was frequently seen attached to the cytoplasmic projections or apical cell membrane of the C-cells. Since the cysts progressively increase in volume and number with age, it is suggested that they may partly play a role in the storage of excess or unneeded hormonal products.
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  • 35
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    Cell & tissue research 245 (1986), S. 91-100 
    ISSN: 1432-0878
    Keywords: Testis ; Leydig cells ; Crystalloids ; Ultrastructure ; Rat, Rattus fusdpes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of Leydig cells in a seasonally breeding rodent, Rattus fuscipes, was studied in the breeding and non-breeding season and compared with Leydig cell morphology after suppression of gonadotrophin secretion induced by hypophysectomy or chronic administration of testosterone. Serum luteinizing hormone (LH) and testosterone (T) were measured and in-vitro T production by testes was assessed by stimulation with human chorionic gonadotrophin (hCG). In non-breeding wild-trapped rats and rats with experimental suppression of gonadotrophins, the Leydig cells were atrophied and exhibited variable amounts of cytoplasmic lipid and crystalloid inclusions, the latter commonly dominating the cytoplasmic area. Compared with fertile rats, serum LH and hCG-stimulated T production of experimentally regressed rats was significantly reduced, confirming structural features indicative of Leydig cell inactivity. Atrophy of Leydig cell nuclei was accompanied by the formation of unusual intranuclear vesicles sometimes containing small crystalloids. Ultrastructural analysis suggested transfer of the vesicles to the cytoplasm where their unification gave rise to much larger crystalloid bodies. Crystalloids occurred when serum LH was depressed and with either full (T treatment) or arrested spermatogenesis (hypophysectomy) suggesting that their formation is governed by pituitary function and is not dependent upon the degree of spermatogenic activity.
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  • 36
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    Cell & tissue research 246 (1986), S. 607-621 
    ISSN: 1432-0878
    Keywords: Rod photoreceptor ; Light damage ; Albino rat ; Ultrastructure ; Reversible change
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The temporal sequence of ultrastructural changes induced in the rat rod photoreceptor by 80 lux light-stress has been studied. The changes seen were compared with those produced by a much dimmer (3 lux) illumination. Some of the early signs of abnormality were (1) degradation of some disk membranes at the tips of outer segments, (2) disaggregation and detachment of ribosomes, (3) lighter matrices in swollen mitochondria, (4) disappearance of the Golgi apparatus, (5) proliferation of autophagic bodies in the inner segments, and (6) appearance of perimitochondrial membrane whorls in the synaptic terminals. No single change could be identified that would inexorably lead to cell death. The overall picture, however, suggested that an inability of the cell to maintain its anabolic balance is responsible for the pyknosis that occurs when the 80 lux exposure exceeds 12–15 h. All changes were reversible when exposure duration did not exceed 12 h, the normal length of the light cycle for these rats.
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  • 37
    ISSN: 1432-0878
    Keywords: Smooth-muscle cell ; Cell culture ; Ultrastructure ; Optical measurement ; Cell contraction ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In numerous investigations using cultured smooth-muscle cells, investigators have consistently added 10–20% fetal calf serum (FCS) to the medium to maintain viable cells. In the present study we utilized an optical technique to investigate whether smooth-muscle cells, cultured with or without FCS, maintain their contractile activity in vitro. With such optical measurement, we were able to detect signals due to spontaneous contractions, in muscle cells cultured in FCS-free medium for up to 8 days, and, for the first time, were also able to observe the conduction of these cell contractions. The ultrastructural characteristics of cultured smooth-muscle cells during contractile activity, were also examined by electron microscopy. The cells were mature and well-differentiated, and were packed with numerous myofilaments. They had developed long cell processes, and were linked to one another by gap junctions. These observations indicated that the smooth-muscle cells, cultured without FCS for 7 to 8 days, were morphologically mature and maintained their contractile activity, whereas the cells cultured in FCS-containing medium showed no detectable signs of contractile activity.
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  • 38
    ISSN: 1432-0878
    Keywords: Chemoreceptor ; Ultrastructure ; Parasitic copepod crustacean ; Pachypygus gibber
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The topography, external structure and ultrastructure of a cephalic sense organ, described for the first time, were studied by light and electron microscopy in the parasitic copepod Pachypygus gibber. This species is unusual in that it has three reproductive sexual forms (two males, one female).The cephalic organ, present only in the atypical male, is made up of numerous functional units, each composed of 4 cells: two sensory cells, one basal enveloping cell and one apical canal-forming cell opening outside via a pore. Many hundred pores are situated within the cuticle of the ventral pleural borders. Through each pore protrude two ciliary endings. An interesting feature is that the ciliary ends are without cuticular cover and thus, directly exposed to the surroundings, a situation unique in arthropods. The structural characteristics of this sense organ and the particular mode of life of the atypical male (with an additional free planktonic phase), lead to the hypothesis that its function is linked to chemical reception in the complex behavioural patterns such as host and sex recognition, during the free life.
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  • 39
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    Cell & tissue research 245 (1986), S. 457-460 
    ISSN: 1432-0878
    Keywords: Anterior pituitary ; Somatotroph ; Lactotroph ; Ultrastructure ; Immunocytochemistry ; Fetal and neonatal rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of immunoreactive somatotrophs and lactotrophs in pituitaries of fetal rats at 19, 20 and 21 days of gestation and on the day of birth was studied. Somatotrophs, first detectable at 19 days of gestation, undergo only minor modifications before reaching the structure described for adults. In particular there is an increase in the endoplasmic reticulum and Golgi apparatus. Lactotrophs, first identifiable in newborn rats, are very different in ultrastructure from adult cells, because the secretory granules are generally small, but variable in shape and size, and the Golgi complex is prominent.
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  • 40
    ISSN: 1432-0878
    Keywords: Organ culture ; Amelogenesis ; Dentinogenesis ; Ultrastructure ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Molar tooth germs from three-day-old rats were cultured successfully for fourteen days, permitting the study of the development in vitro of both extracellular matrix and cellular elements such as odontoblasts and ameloblasts. The ultrastructure of the cultured tooth germs was compared with the ultrastructure of tooth germs in vivo at a comparable developmental stage. Progenitor cells of odontoblasts and ameloblasts were found to differentiate in vitro. Odontoblasts seemed to contain more lysosome-like bodies and fewer secretory granules than in vivo. They formed normally mineralizing dentine or a thick layer of dense, unmineralized predentine with incidentally some amorphous, extracellular material. Enamel was exclusively present opposite well developed dentine. It was often hyperor hypomineralized and enamel rods were not as regularly shaped as in vivo. In places where no enamel formation had taken place, large amounts of amorphous extracellular material were sometimes seen. From these observations it can be concluded that cellular development in cultured tooth germs appeared more or less normal, but extracellular matrix formation and mineralization were sometimes disturbed.
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  • 41
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    Cell & tissue research 244 (1986), S. 165-171 
    ISSN: 1432-0878
    Keywords: Iridophores ; Motility ; Innervation ; Ultrastructure ; Teleost (Odontobutis obscura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Reflecting chromatophores in the dermis of the skin of a freshwater goby, Odontobutis obscura, are of an iridophore type. These chromatophores contain numerous reflecting platelets, which are similar to those in iridophores of other fish and amphibian species. It was found that these iridophores are motile, i.e., these cells respond to certain stimuli with translocation of the platelets within the cells. K+ ions induced dispersion of the platelets in excised scale preparations, but not in excised scales from chemically denervated fish. Norepinephrine and melatonin also induced dispersion of the platelets. Alpha-MSH was effective in aggregating these organelles into the centrospheres of the cells. The conclusions reached are: (1) iridophores of O. obscura are motile; (2) the movement of the iridophores is under nervous and hormonal control.
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  • 42
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    Cell & tissue research 243 (1986), S. 405-414 
    ISSN: 1432-0878
    Keywords: Leydig cells ; Ethane dimethanesulphonate ; Ultrastructure ; Destruction ; Gonadotrophins ; Testosterone ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The cytotoxic effects of ethane dimethanesulphonate upon rat Leydig cells were examined ultrastructurally up to 3 days after treatment and related to changes in serum levels of gonadotrophins and testosterone. Six hours after administration of ethane dimethanesulphonate the usual tubulo-vesicular morphology of Leydig-cell smooth endoplasmic reticulum was converted to small vesicles and the Golgi apparatus showed focal hypertrophy into anastomosing tubules. These changes became more marked by 12 h with many Leydig cells exhibiting karyopyknosis and hyperchromatism. Necrotic Leydig cells were often engulfed by macrophages, the latter containing pyknotic fragments of Leydig cells within their cytoplasm. One day after administration, advanced necrosis of Leydig cells occurred, many of which were phagocytosed by macrophages, and on day 3, destruction of Leydig cells was complete resulting in their elimination from the interstitial tissue, which contained only loose connective tissue and macrophages. Structural alterations to the Leydig cells from 6–24 h was reflected by a significant reduction in serum testosterone levels which further declined to the limits of detection accompanying the abolition of Leydig cells on day 3. These changes were paralleled by a significant elevation of serum LH and FSH levels suggesting diminished feedback regulation of pituitary gonadotrophin secretion. The results indicate that ethane dimethanesulphonate is a rapidly acting Leydig cell toxin which may be a useful experimental tool in further studies of spermatogenic function mediated via Sertoli cell-Leydig cell interaction.
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  • 43
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    Cell & tissue research 244 (1986), S. 493-499 
    ISSN: 1432-0878
    Keywords: Endothelium ; Capillaries ; Kidney ; Ultrastructure ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A new endothelial cell structure, named the endothelial pocket, has been found by combined transmission and scanning electron microscopic studies of renal peritubular capillaries. Transmission EM observations made on these and other fenestrated capillaries demonstrated that each pocket consists of an attenuated fold of fenestrated endothelium that projects 200 nm into the lumen above the rest of the endothelial surface. Beneath this luminal fold, there is a space and then another layer of fenestrated endothelium which abuts the basal lamina. The linear density of endothelial pockets was measured in the capillaries of the kidney cortex, intestinal mucosa and exocrine pancreas in mice and determined to be 0.067, 0.017 and 0.007 pockets·μm-1 respectively. Cationic ferritin decoration of the anionic sites on the luminal surface of the endothelium in these capillary beds revealed that both unlabelled and labelled diaphragms are clustered. In such specimens, the majority of the luminal diaphragms on endothelial pockets did not have cationic ferritin binding sites detectable by either scanning or transmission EM. On this account as well as on account of their general morphology, endothelial pockets appear to be multifold versions of the simple transendothelial channel.
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  • 44
    ISSN: 1432-0878
    Keywords: Capillary ; Epithelium ; Retina ; Retinal pigment epithelium ; Ultrastructure ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Light- and urethane-induced retinopathies in rats are characterized by loss of photoreceptors. Retinal capillaries subsequently become incorporated into the normally avascular retinal pigment epithelium. These models provided an opportunity to study the response of epithelial cells to closely apposed capillaries, in order to determine if capillaries contribute to the polar organization of epithelial cells. Pigment epithelial cells reorganized their lateral plasma membrane where the latter faced intraepithelial capillaries. This normally flat, undifferentiated membrane developed attachment sites, folds and intracytoplasmic tubules, and exhibited endocytosis and putative basal lamina secretion. These structural and functional specializations are normally restricted to the basal plasma membrane — the normal vascular front of the cell facing the dense meshwork of capillaries constituting the choriocapillaris. We conclude that RPE cells, and perhaps epithelia in general, polarize in response to an adjacent capillary bed.
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  • 45
    ISSN: 1432-0878
    Keywords: VIP ; Nerve fibers ; Ganglion cells ; Chromaffin cells ; Ultrastructure ; Adrenal gland ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary VIP-like immunoreactivity was revealed in a few chromaffin cells, medullary ganglion cells and a plexus of varicose nerve fibers in the superficial cortex and single varicose fibers in the juxtamedullary cortex and the medulla of the rat adrenal gland. VIP-like immunoreactive chromaffin cells were polygonal in shape without any distinct cytoplasmic processes and they appeared solitarily. Their cytoplasm contained abundant granular vesicles having a round core and the immunoreactive material was localized to the granular core. VIP-immunoreactive ganglion cells were multipolar and had large intracytoplasmic vacuoles. The immunoreactive material was localized not only in a few granular vesicles but also diffusely throughout the axoplasm. VIP-immunoreactive varicose nerve fibers in the superficial cortex were characterized by abundant small clear vesicles and some large granular vesicles, while those in the juxtamedullary cortex and medulla and the ganglionic processes were characterized by abundant large clear vesicles, as well as the same vesicular elements as contained in the nerves in the superficial cortex. The immunoreactive material was localized on the granular cores and diffusely in the axoplasm in both nerves. Based on the similarity and difference in the composition of the vesicles contained in individual nerves, it is likely that the VIP-immunoreactive nerve fibers in the medulla and the juxtamedullary cortex are derived from the medullary VIP-ganglion cells, while those in the superficial cortex are of extrinsic origin. The immunoreactive nerve fibers in both the cortex and the medulla were often in direct contact with cortical cells and chromaffin cells, where no membrane specializations were formed. The immunoreactive nerve fibers were sometimes associated with the smooth muscle cells and pericytes of small blood vessels in the superficial cortex. In addition they were often seen in close apposition to the fenestrated endothelial cells in the cortex and the medulla, only a common basal lamina intervening. Several possible mechanisms by which VIP may exert its effect in the adrenal gland are discussed.
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  • 46
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    Cell & tissue research 243 (1986), S. 137-144 
    ISSN: 1432-0878
    Keywords: Lung ; Blood-air barrier ; Surfactant ; Ultrastructure ; Freeze fracture ; Birds ; Penguin (Aptenodytes forsten)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Of all avian species the emperor penguin is the best adapted bird to attain the greatest diving depths and diving durations. Therefore the lung of this bird was investigated with electron-microscopic, i.e., freeze-fracture and thin-section methods. The parabronchi are surrounded by bundles of smooth muscle cells innervated by varicosities of autonomic nerves. The parabronchial epithelium is flat, bears a few microvilli and does not show any conspicuous ultrastructural specializations; only individual cells contain secretory granules. The atrial epithelial cells bear apical microvilli and are interconnected by adhering and tight junctions (5–10 sealing strands), the latter presumably forming an effective barrier against paracellular fluid movements. The cells contain lamellar inclusions of two types: (i) round membrane-bounded granules, the lamellar content of which is fixation-labile, and (ii) large polymorphic compact deposits of well-preserved lamellae. In both types of inclusions the individual lamellae can be of trilaminar appearance, whereas their fracture faces are smooth. Lamellar material also covers the epithelium of atria, infundibula and air capillaries. In thin areas the diameter of the morphological blood-air barrier measures 220–330 nm. Usually the endothelium of the blood capillaries is thicker (40–180 nm) than the air capillary epithelium (25–150 nm). Both epithelium and endothelium are interconnected by tight junctions, which seem to be more extensive and presumably tighter in the epithelium than in the endothelium. Frequently the common basal lamina is the thickest individual component of the blood-air barrier, measuring between 170–230 nm. Often collagen fibrils occur in this area of the barrier. In comparison with that of other birds the entire blood-air barrier of the emperor penguin is relatively thick, probably owing to an adaptation of the lung tissue which must resist high hydrostatic pressure during diving excursions.
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  • 47
    ISSN: 1432-0878
    Keywords: Parathyroid gland ; Protein A-gold technique ; Ultrastructure ; Storage granules ; Mammals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Electron-microscopic immunocytochemistry for the demonstration of parathormone in parathyroid chief cells was performed in adult male rats, gerbils, mice, and dogs, using the protein A-gold technique. Protein A-gold particles were detected over both large and small secretory granules in all the animals examined. In the former, they were concentrated not only over type-I granules with a large core, but also over type-II granules with a small core. They were also located over atypical granules, including heterogeneously dense granules, granules having vesicles in a finely particulate core, and distorted granules. All labelled secretory granules were characterized by the presence of a clear halo of varying width around the core. Occasionally, Golgi cisternae as well as Golgi vacuoles with a finely particular content were also labelled. The labelling of the secretory granules was strong in dogs, moderate in rats and gerbils, and weak in mice. In addition, it was more intense in the non-osmicated preparations than in the osmicated preparations. The frequency of both types of large granules showed species differences. The possible factors involved in these differences are discussed.
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  • 48
    ISSN: 1432-0878
    Keywords: Peripolar cells ; Juxtaglomerular apparatus ; Kallikrein ; Ultrastructure ; Mammals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Peripolar cells are granulated glomerular epithelial cells that form a cuff around the vascular pole of the glomerulus. Quantitation of these cells in 17 species of mammals (including man, several laboratory animals and a variety of other species) indicated that they were detectable by light microscopy in all but one of the mammals that were examined (the Australian hopping mouse). In adult mammals with detectable peripolar cells, the “peripolar cell index” (the percentage of randomly sectioned glomeruli that displayed peripolar cells in histological sections of kidney) ranged from 0.15 (for echidna) to 11.86 (for sheep). Newborn lambs and rats showed strikingly high values (23.30 and 10.76, respectively) compared with their adult counterparts. Using electron microscopy, peripolar cells were observed in all species that were examined, including the Australian hopping mouse. Morphologically, peripolar cells were similar in all species although their size and granule population varied. They showed a predominantly outer cortical glomerular distribution and a close anatomical relationship with the renin-containing myoepithelioid cells. These findings indicate that peripolar cells are present in a wide variety of species and support the view that such cells may play a significant role in the regulation of normal renal function.
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  • 49
    ISSN: 1432-0878
    Keywords: Uterus ; Epithelium ; Ultrastructure ; Pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of the endometrial epithelium of the pig was studied during the estrous cycle and early pregnancy up to implantation. Special attention was given to the luminal epithelium and morphological indications of protein synthesis. Although the general morphology of the luminal and glandular epithelia is similar (both tissues consist of secretory cells and ciliated cells at all the stages studied), it appears that the two epithelia should be considered as two functionally different units in the pre-implantation period. Morphological evidence suggests the presence of at least three different secretory products within luminal epithelial cells; they are released at different times, i.e. at estrus, between day 8 and 10 and after day 11. The glandular epithelium shows release of secretory products from day 10–11. Increasing amounts of glycogen were found within epithelial cells, especially in pregnant gilts from day 12. The possible significance of secretory activity of the epithelium is discussed in relation to the development of the embryos.
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  • 50
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    Plant systematics and evolution 153 (1986), S. 259-264 
    ISSN: 1615-6110
    Keywords: Gymnosperms ; Angiosperms ; Pteridosperms ; Caytoniales ; Caytonanthus ; Ultrastructure ; pollen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The pollen grain wall structure ofCaytonanthus arberi was studied with SEM and TEM. The monosulcate, bisaccate pollen grain of this taxon has an alveolar wall structure that is more like that found in saccate Conifers than primitive Angiosperms. Pollen grain morphology does not support a Caytonialean ancestry for the Angiosperms.
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  • 51
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    Plant systematics and evolution 154 (1986), S. 31-39 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Lactoridaceae ; Ultrastructure ; pollen ; fossil pollen (Cretaceous)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The pollen wall ultrastructure of the primitive AngiospermLactoris fernandeziana Phil. (Lactoridaceae) is described. The monosulcate aperture, granular wall structure and sacci (all primitive features) suggest placement of this family in theMagnoliales. Pollen ofLactoris is compared to fossil dispersed pollen from the Lower and lower-Upper Cretaceous. The fossil pollen shares characteristics which are restricted to theLactoridaceae suggesting that this family was present during the early Cretaceous.
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  • 52
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    Protoplasma 131 (1986), S. 1-10 
    ISSN: 1615-6102
    Keywords: Cell division ; Lomentaria ; mitosis ; Red algae ; Rhodophyta ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mitosis in the marine red algaLomentaria baileyana (Rhodymeniales, Rhodophyta) was studied with the electron microscope. Nucleus associated organelles known as polar rings (PRs) migrate to establish the division poles at prophase. At prometaphase, shallow invaginations in the nuclear envelope (NE) form on two sides of each PR and soon rupture. The gaps that are consequently formed contain several small fragments of NE. A larger region of NE remains intact between the two gaps. By metaphase several cisternae of perinuclear endoplasmic reticulum (PER) have enclosed most of the nucleus but remain absent from the polar regions. The nucleolus disperses partially and a typical metaphase plate of chromosomes is formed. Each PR has disjoined into separate proximal and distal portions. MTs converge widely on all regions of the polar area, but do not extend into the cytoplasm. Some MTs end near or at the chromosomes while others extend slightly farther past the chromosomes or diagonally to the NE. As chromosomes move to opposite poles at anaphase, they are accompanied by nucleolar material. An interzonal midpiece (IZM) is created as the pole to pole distance increases and the NE remains intact except for the polar gaps. Following detachment from the IZM, the daughter nuclei are separated by a large central vacuole as a cleavage furrow develops and eventually constricts to form two cells following pit connection formation. It is suggested that mitosis inLomentaria represents an evolutionary intermediate between that seen in the higher and lower groups of red algae. This conclusion is in agreement with conventional morphological and light microscopic criteria used to placeLomentaria in theRhodymeniales, which is considered to be the next to most advanced order in theRhodophyta.
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  • 53
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    Protoplasma 134 (1986), S. 154-162 
    ISSN: 1615-6102
    Keywords: Cell division ; Leishmania ; Mitosis ; Ultrastructure ; Kinetoplastida
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The interphase nucleus ofLeishmania adleri has clumps of chromatin associated with the nuclear envelope and a large centrally located nucleolus. Prior to mitosis the basal bodies replicate at the cell anterior. Subsequently, dense plaques appear in the equatorial region of the nucleus at the time of spindle development. Microtubules appear in the nucleus adjacent to the nuclear envelope and embedded in the matrix of the plaques. A central spindle composed of a single bundle of microtubules develops and spans the nucleus. Plaques and nucleolar components laterally associate with the spindle and migrate towards the poles. The central spindle elongates to three to four times its original length separating the forming daughter nuclei and producing an interzonal spindle. A remnant of the interzonal spindle remains attached to each of the daughter nuclei until late into cytokinesis. The kinetoplast does not divide until after the completion of mitosis.
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  • 54
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    Protoplasma 131 (1986), S. 166-173 
    ISSN: 1615-6102
    Keywords: Chloroplast development ; Pisum sativum ; Prolamellar body ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We analyzed transformation, recrystallization, splitting and dispersion of prolamellar bodies during chloroplast development in pea seedlings illuminated by white, red and blue light of low intensity. With the help of a stereometric method we determined that there was a significant increase of prolamellar body number and a sharp decrease of their volume in differentiating chloroplast even in the first 2 hours of illumination. Decrease of prolamellar body dimensions was due both to gradual dispersion of its elements into primary thylakoids (indicated by the decrease of total volume of prolamellar bodies in plastid) and to splitting of prolamellar bodies (indicated by the increase of number of promellar bodies in plastid). Red light was more effective in transformation, splitting and dispersion of prolamellar bodies than blue light during the first 8–12 hours. Longer treatment with blue light had a stronger influence on these processes and on complete recrystallization than other light treatments.
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  • 55
    ISSN: 1615-6102
    Keywords: Chlamydomonas ; Endoplasmic reticulum ; Golgi apparatus ; Wall mutants ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Ultrastructural observations on the principal endomembranes (endoplasmic reticulum, Golgi apparatus) of synchronously growing of wild type and mutant (CW 2, CW 15) strains ofChlamydomonas reinhardii have been carried out. The dictyosomes of the Golgi apparatus in all three cases are highly polar in morphology but lack intercisternal filaments. A clear spatial relationship between dictyosomes and endoplasmic reticulum is seen and a transfer of vesicles from the latter to the former is easily visualized. Coated vesicles invariably appear to be restricted to the trans-pole of the dictyosomes. The endoplasmic reticulum adjacent to the cis pole of dictyosomes is considerably hypertrophied in the case of the wild type, only partially so in the mutant CW 2 but not at all in the mutant CW 15. In the wild type this swelling is most extreme during the period of wall deposition and for several hours afterwards. The results are discussed in relation to the biosynthesis and intracellular transport of, particularly O-glycosidically linked, glycoproteins.
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  • 56
    ISSN: 1615-6102
    Keywords: Chloroplast development ; Grana formation ; Pisum sativum ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We analyzed the formation of thylakoids and grana during the development of pea chloroplasts, illuminated by white, red and blue low intensity light. The total length of granal and intergranal thylakoids, and the length of granal thylakoids per unit area of plastid section were measured. Initially the greatest increase in length of granal thylakoids and the highest incidence of grana with large thylakoid content occurred in red light. On the other hand, with illumination times of over 12 hours blue light appeared to be more efficient in stimulating grana formation and thylakoid growth.
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  • 57
    ISSN: 1615-6102
    Keywords: Diapensia lapponica ; Mitochondria ; Morphometry ; Seasonal variation ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Seasonal changes in the mitochondrial ultrastructure were examined in palisade parenchyma cells of a tuft-formingDiapensia lapponica L. collected at monthly intervals in Northern Finland. Quantitative analyses to measure volume and surface densities were conducted during different periods of growth (stages of growth, acclimation, winter period and deacclimation) in the annual cycle. The volume density was highest in the summer and lowest in the spring; the difference was significant with both fixatives used GA and GA/FA. The largest membrane area (the mitochondrial outer membrane and the cristal membranes together) was observed in the summer and autumn, and was significantly less in the winter and spring. This correlated with fewer mitochondria in the spring and a smaller number of cristae in the winter and spring. In the material fixed in GA/FA the distribution of length/width ratios of mitochondria was relatively uniform in all seasons. However, the mitochondrial ultrastructure had the most varied appearance during the winter. Hypertonie GA/FA solution did not cause significant differences either in the ultrastructure or the volume and surface densities of the mitochondria.
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  • 58
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    Protoplasma 135 (1986), S. 102-111 
    ISSN: 1615-6102
    Keywords: Ultrastructure ; Haptoglossa mirabilis ; Differentiation ; Gun cell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The most prominent ultrastructural characteristics of the cyst ofHaptoglossa mirabilis are a large centrally-placed nucleus which is partially ringed by three or four parallel cisternae of rough endoplasmic reticulum (r-ER), a centriole pair and single large Golgi complex which occupy the anterior end of the cell, and a population of provacuoles which occupies the posterior. During germination these organelles migrate into a narrow germ tube which subsequently expands to form the gun cell initial. The extracellular components of the attack apparatus (i.e. missile and injection tube) are formed entirely in the developing gun cell; indirect evidence suggests that both the Golgi complex and r-ER are involved in their synthesis. The intra-cellular component of the attack apparatus comprises the posterior, anterior and apical vacuoles. The posterior vacuole forms by fusion and expansion of the original cyst provacuoles; the formation of the anterior and apical vacuoles occurs late in gun cell differentiation and involves fusion of Golgi-derived vesicles.
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  • 59
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    Protoplasma 134 (1986), S. 102-110 
    ISSN: 1615-6102
    Keywords: Isoelectric focusing ; Isolation ; Potato tuber cell walls ; Purification ; Salt extraction ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A preparative procedure for cell wall isolation and purification was developed. The purity of the isolated cell walls was judged biochemically by the lack of activity of cytoplasmic marker enzymes and morphologically by examination at both the light and electron microscope levels. The purified cell walls were extracted with various salt treatments and the molecular weight range of most of the extracted proteins was between 14 and 31 kDa. The salt extracted hydrolytic enzymes were basic in nature (pI〉7.0) compared to their cytosolic counterparts (pI〈7.0). Some enzymes were readily extracted from cell walls (β-glucosidase and β-NAcglucosaminidase) with high salt treatment while most of the α-mannosidase activity associated with purified cell walls could not be removed even with sequential high salt treatments.
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  • 60
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    Protoplasma 135 (1986), S. 119-129 
    ISSN: 1615-6102
    Keywords: Differentiation ; Pectin ; Phytophthora ; Surface ; Ultrastructure ; Zoospore
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Populations ofPhytophthora palmivora zoospores induced to undergo synchronous differentiation by the addition of pectin showed an ordered sequence of surface changes. Within 10 seconds “blebs” and “microvilli” appeared on the cell and flagellar surfaces. These projections gave the zoospore a highly irregular and even convoluted surface for up to 30 seconds poststimulation. Between 30 and 40 seconds, pits appeared on the surface, which began to assume a smoother texture. Zoospores then rounded up and by 4 minutes had assumed an almost spherical form. At this time the surface became wrinkled as though shrinkage had occurred. Germ tubes could be seen emerging from almost all cells by 40 minutes, and as germ tubes enlarged, the cell surface became smooth again but with a fibrous texture. Many zoospores retained their flagella through to the germling stage. When cells were stimulated in iso-osmotic rather than hypo-osmotic media, germ tubes in most cases emerged at or very near the site of flagellar attachment. Differentiation in iso-osmotic media showed the same sequence except that the cells did not assume the spherical shape normally characteristic of cysts and germlings differentiating under hypo-osmotic conditions. Strontium-induced differentiation again followed the same general sequence as desribed above, although the cell surface was noticeably more convoluted and irregular prior to the stage of the formation of pits, and flagella were always shed between 40 seconds and 1 minute. Calcium addition up to 20 seconds after the initiation of differentiation with pectin prevented a high proportion of zoospores from encysting, the cells remained motile, and although the frequency of surface blebs and microvilli increased, pits did not appear. These surface changes are consistent with those expected during stimulus-mediated secretion, and it appears that calcium is able to inhibit differentiation only before the secretory step (pit formation).
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  • 61
    ISSN: 1615-6102
    Keywords: Dark-stress ; Recovery ; Root-nodules ; Soybean ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Root-nodules of soybean plants dark-stressed for 8 days and then allowed to recover for up to 17 days were examined by transmission electron microscopy. Control nodules possessed all the ultrastructural features characteristic of infected and uninfected nodule cells. Minimal changes in the appearance of host cells and bacteroids occurred during the first four days of dark stress. After 8 days of dark stress, damage was observed in the cellular and organelle membranes; however, very few changes were observed in the bacteroids. Nodule structure continued to degrade during the first two days of recovery after which time nodules either recovered or completely degraded. In the former case, structural integrity returned to all nodule cells. In the latter case all structural integrity of the host cell disappeared; however, bacteroids appeared intact suggesting that they remained viable.
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  • 62
    ISSN: 1615-6102
    Keywords: Root meristems ; Cell plate ; Ultrastructure ; 2,6-dichlorobenzonitrile
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The effect of 2,6-dichlorobenzonitrile on cytokinesis of meristematic cells of onion root during both treatment and recovery has been studied by electron microscopic techniques. 2,6-dichlorobenzonitrile interferes with cell plate formation in such a way that Golgi apparatus vesicles of treated cells appear to be different than controls and seem to coalesce as anomalous partial cell plates. During recovery, an apparently normal progression of cytokinesis is observed and abnormal portions of the cell plate are retained. Nuclear constrictions are observed frequently during recovery as a result of temporal alterations in cytokinesis. Our results show that 2,6-dichlorobenzo-nitrile induces anomalous and/or incomplete cell plates, which might be caused by an altered function of Golgi apparatus.
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  • 63
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    European journal of plastic surgery 9 (1986), S. 122-129 
    ISSN: 1435-0130
    Keywords: Congenital lymphoedema in dogs ; Ultrastructure ; Chronic lymphoedema ; Chronic inflammation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In two congenitally lymphoedematous dogs, a quantitative fine structural study was made of the subcutaneous tissue. It was found that the blood vessels, inital lymphatics and tissue channels all increased considerably, as did the small vesicles and vacuoles in the endothelium of both blood and lymph vessels. Collagen, fibroblasts, macrophages, and fat containing macrophages “fat cells” all increased greatly, while the small and medium lymphocytes increased to a moderate extent. These results are similar to those found in chronic experimental lymphoedema in dogs, in chronic high-protein oedema and in chronic inflammation. The underlying similarity of all these conditions is emphasised.
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    European archives of oto-rhino-laryngology and head & neck 243 (1986), S. 265-268 
    ISSN: 1434-4726
    Keywords: Respiratory cilia ; Measurement of ciliary orientation ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We have created a method for measuring the orientation of cilia in the respiratory epithelium. Ciliary orientation is the direction perpendicular to the plane defined by the central tubules of the cilia and is an estimate of ciliary beat direction. Ciliary orientation can be estimated by measuring the angle between the plane defined by the central tubules and a reference line. The standard deviation of these measurements describes the variation present in the beat directions of the cilia. The reference line must be so chosen that the majority of measurements falls at about the middle of the 0°–180° range. We tested measurements by using both a glass angle measure and a semiautomatic image analyzer (IBAS I). The latter approach was faster and more reproducible. We made our measurements of normal tissues on samples obtained from two healthy adult non-smokers. Measurements were made in four areas of each sample, with 59–110 cilia in each. The differences between the maximum and minimum angles of the ciliary orientation in the same area varied from 167.9° to 85.4° from the reference line. The standard deviation varied from 18.0° to 35.4° and we consider this to be a normal variation in ciliary orientation. Of the cilia, 57% were within a standard deviation of 20°.
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  • 65
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    Proteins: Structure, Function, and Genetics 1 (1986) 
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
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  • 66
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 1-1 
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
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  • 67
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 2-3 
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
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  • 68
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 4-15 
    ISSN: 0887-3585
    Keywords: membrane biomimetic system ; reverse micelles ; interfacial water ; myelin proteins ; solid enzyme activity ; organic solvents ; biotechnology ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Additional Material: 11 Ill.
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 16-22 
    ISSN: 0887-3585
    Keywords: protein design ; alpha-helical bundle ; x-ray crystallography ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Twelve- and sixteen-residue peptides have been designed to form tetrameric alphahelical bundles. Both peptides are capable of folding into amphiphilic alpha-helices, with leucyl residues along one face and glutamyl and lysyl residues along the opposite face. Four such amphiphilic alpha-helices are capable of forming a noncovalently bonded tetramer. Neighboring helices run in antiparallel directions in the design, so that the complex has 222 symmetry. In the designed tetramer, the leucyl side chains interdigitate in the center in a hydrophobic interaction, and charged side chains are exposed to the solvent. The designed 12-mer(ALPHA-1) has been synthesized, and it forms helical aggregates in aqueous solution as judged by circular dichroic spectroscopy. It has also been crystallized and characterized by x-ray diffraction. The crystal symmetry is compatible with (but does not prove) the design. The design can be extended to a four-alpha-helical bundle formed from a single polypeptide by adding three peptide linkers.
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  • 70
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 23-33 
    ISSN: 0887-3585
    Keywords: peptide helix ; protein stability ; framework model of folding ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Recent work has shown that, with synthetic analogues of C-peptide (residues 1-13 of ribonuclease A), the stability of the peptide helix in H2O depends strongly on the charge on the N-terminal residue. We have asked whether, in semisynthetic ribonuclease S reconstituted from S-protein plus an analogue of S-peptide (1-15), the stability of the peptide helix is correlated with the Tm of the reconstituted ribonuclease S. Six peptides have been made, which contain Glu9 → Leu, a blocked α-COO- group (—CONH2), and either Gln11 or Glu11. The N-terminal residue has been varied; its charge varies from +2 (Lys) to -1 (succinyl-Ala). We have measured the stability of the peptide helix, the affinity of the peptide for S-protein (by C.D. titration), and the thermal stability of the reconstituted ribonuclease S.All six peptide analogues show strongly enhanced helix formation compared to either S-peptide (1-15) or (1-19), and the helix content increases as the charge on the N-terminal residue changes from +2 to -1. All six peptides show increased affinity for S-protein compared to S-peptide (1-19), and all six reconstituted ribonucleases S show an increase in Tm compared to the protein with S-peptide (1-19). The Tm increases as the charge on residue 1 changes from +2 to -1. The largest increment in Tm is 6°.The results suggest that the stability of a protein can be increased by enhancing the stability of its secondary structure.
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 43-46 
    ISSN: 0887-3585
    Keywords: protein stability ; helix-coil ; mutant ; calorimetry ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Oligonucleotide-directed mutagenesis has been used to replace α-helical glycines in the N-terminal domain of λ repressor with alanines. Since alanine is a significantly better helix-forming residue than glycine, these changes were predicted to have a stabilizing effect. We show that the Gly46→Ala substitution, the Gly48→Ala substitution, and the double substitution increase the melting temperature of the N-terminal domain by 3-6°.
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  • 72
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 34-42 
    ISSN: 0887-3585
    Keywords: hydrogen exchange ; BPTI ; folding pathway ; protein dynamics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A method to be used for experimental studies of protein folding introduced by Schmid and Baldwin (J. Mol. Biol. 135: 199-215, 1979), which is based on the competition between amide hydrogen exchange and protein refolding, was extended by using rapid mixing techniques and 1H NMR to provide site-resolved kinetic information on the early phases of protein structure acquisition. In this method, a protonated solution of the unfolded protein is rapidly mixed with a deuterated buffer solution at conditions assuring protein refolding in the mixture. This simultaneously initates the exchange of unprotected amide protons with solvent deuterium and the refolding of protein segments which can protect amide groups from further exchange. After variable reaction times the amide proton exchange is quenched while folding to the native form continues to completion. By using 1H NMR, the extent of exchange at individual amide sites is then measured in the refolded protein. Competition experiments at variable reaction times or variable pH indicate the time at which each amide group is protected in the refolding process. This technique was applied to the basic pancreatic trypsin inhibitor, for which sequence-specific assignments of the amide proton NMR lines had previously been obtained. For eight individual amide protons located in the β-sheet and the C-terminal α-helix of this protein, apparent refolding rates in the range from 15s-1 to 60 s-1 were observed. These rates are on the time scale of the fast folding phase observed with optical probes.
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 47-59 
    ISSN: 0887-3585
    Keywords: protein electrostatics ; substrate diffusion ; Poisson-Boltzmann ; electrostatic potentials ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: In this paper we report the implementation of a finite-difference algorithm which solves the linearized Poisson-Boltzmann equation for molecules of arbitrary shape and charge distribution and which includes the screening effects of electrolytes. The microcoding of the algorithm on an ST-100 array processor allows us to obtain electrostatic potential maps in and around a protein, including the effects of ionic strength, in about 30 minutes. We have applied the algorithm to a dimer of the protein Cu-Zn superoxide dismutase (SOD) and compared our results to those obtained from uniform dielectric models based on coulombic potentials. We find that both the shape of the protein-solvent boundary and the ionic strength of the solvent have a profound effect on the potentials in the solvent. For the case of SOD, the cluster of positive charge at the bottom of the active site channel produces a strongly enhanced positive potential due to the focusing of field lines in the channel - a result that cannot be obtained with any uniform dielectric model. The remainder of the protein is surrounded by a weak negative potential. The electrostatic potential of the enzyme seems designed to provide a large cross-sectional area for productive collisions. Based on the ionic strength dependence of the size of the positive potential region emanating from the active site and the repulsive negative potential barrier surrounding the protein, we are able to suggest an explanation for the ionic strength dependence of the activity of the native and chemically modified forms of the enzyme.
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 66-73 
    ISSN: 0887-3585
    Keywords: protein domain ; polymerase ; 3′-5′ exonuclease ; artificial gene ; expression vector ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The Klenow fragment of DNA polymerase I from Escherichia coli has two enzymatic activities: DNA polymerase and 3′-5′ exonuclease. The crystal structure showed that the fragment is folded into two distinct domains. The smaller domain has a binding site for deoxynucleoside monophosphate and a divalent metal ion that is thought to identify the 3′-5′ exonuclease active site. The larger C-terminal domain contains a deep cleft that is believed to bind duplex DNA. Several lines of evidence suggested that the large domain also contains the polymerase active site. To test this hypothesis, we have cloned the DNA coding for the large domain into an expression system and purified the protein product. We find that the C-terminal domain has polymerase activity (albeit at a lower specific activity than the native Klenow fragment) but no measurable 3′-5′ exonuclease activity. These data are consistent with the hypothesis that each of the three enzymatic activities of DNA polymerase I from E. coli resides on a separate protein structural domain.
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  • 75
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 60-65 
    ISSN: 0887-3585
    Keywords: computerized data bank ; sequence comparison heuristics ; databank access ; data bank merging ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Four major protein sequence data collections (NBRF-PIR, PSD-Kyoto, PGtrans, and NEWAT) have been merged into a single nonredundant data bank called PseqIP. The data bank entries were automatically matched by a heuristic computer program relying on the fast computation of the number of tetrapeptides shared by two sequences. PseqIP 1.0 includes 6,068 different protein sequences for a total of 1,357,067 residues, representing most of the available sequence information to date. During the course of this work, we found about 600 occurrences course of a protein sequence recorded with a one-amino-acid variation in at least two different data banks. A flat file (ASCII computer-readable format) version of PseqIP 1.0, well-suited for exhaustive homology searches and statistical sequence analysis, is available from our laboratory.
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  • 76
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 74-80 
    ISSN: 0887-3585
    Keywords: antibody ; crystal structure ; anti-galactan ; J539 ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The crystal structure of the Fab of the galactan-binding immunoglobulin J539 (a mouse IgA,κ) has been determined at a resolution of approximately 2.6 Å by X-ray diffraction. The starting model was that obtained from the real space search described previously (Navia, M.A., Segal, D.M., Padlan, E.A., Davies, D.R., Rao, D.N., Rudikoff, S. and Potter, M. “Crystal structure of galactan-binding mouse immunoglobulin J539 Fab at 4.5 Å resolution.” Proc. Nat. Acad. Sci. USA, 76:4071-4074, 1979). This Fab structure has now been refined by restrained least-squares procedures to an R-value of 19% for the 11,690 unique reflections between 8.0 Å and 2.6 Å. The rms deviation from ideal bond lengths is 0.025 Å. The overall structure differs from McPC603 Fab, another mouse IgA,κ antibody, in that the elbow bend, relating the variable and constant parts of the molecule, is 145° vs. 133° for McPC603. The region of the molecule expected to be the antigen binding site contains a large cavity with two clefts leading away from it. This has been fitted with a model of an oligo-galactan.
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  • 77
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 90-99 
    ISSN: 0887-3585
    Keywords: visual excitation ; rhodopsin ; enzyme regulation ; cyclic nucleotide cascade ; G-proteins ; inhibitory subunit ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The switching on of the cGMP phosphodiesterase (PDE) in retinal rod outer segments by activated transducin (Tα-GTP) is a key step invisual excitation. The finding that trypsin activates PDE (αβγ) by degrading its γ subunit and the reversal of this activation by γ led to the proposal that Tα-GTP activates PDE by relieving an inhibitory constraint imposed by γ (Hurley and Stryer: J. Biol. Chem. 257:11094-11099, 1982). We report here studies showing that the addition of γ subunit also reverses the activation of PDE by Tα-GTP-γS. A procedure for preparing γ in high yield (50-80%) is presented. Analyses of SDS polyacrylamide gel slices confirmed that inhibitorya activity resides in the γ subunit. Nanomolar γ blocks the activation of PDE by micromolar Tα-GTPγS. The degree of activation of PDE depends reciprocally on the concentrations of γ and Tα-GTPγS. γ remains bound to the disk membrane during the activation of PDE by transducin. The binding of γ to the αβ subunits of native PDE is very tight; the dissociation constant is less than 10 pM, indicating that fewer than 1 in 1,700 PDE molecules in rod outer segments are activated in the absence of Tα-GTP.
    Additional Material: 11 Ill.
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  • 78
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 81-89 
    ISSN: 0887-3585
    Keywords: protein stability ; protein denaturation ; denatured state ; structural intermediates ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Eleven mutant forms of staphylococcal nuclease with one or more defined amino acid substitutions have been analyzed by solvent denaturation by using intrinsic fluorescence to follow the denaturation reaction. On the basis of patterns observed in the value of m-the rate of change of log Kapp (the apparent equilibrium constant between the native and denatured states) with denaturant concentration - these proteins can be grouped into two classes. For class I mutants, the value of m with guanidine hydrochloride is less than the wild-type value and is either constant or increases slightly with increasing denaturant; the value of m with urea is also less than wild type but shows a marked increase with increasing denaturant concentration, often approaching but never exceeding the wild-type value. For class II mutants, m is constant and is greater than wild type in both denaturants, with the increase being consistently larger in guanidine hydrochloride than in urea. When double or triple mutants are constructed from members of the same mutant class, the change in m is usually the sum of the changes produced by each mutation in isolation. One plausible explanation for these altered patterns of denaturation is that chain-chain or chain-solvent interactions in the denatured state have been modified - interactions which appear to involve hydrophobic groups.
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  • 79
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    Proteins: Structure, Function, and Genetics 1 (1986) 
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
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  • 80
    ISSN: 0887-3585
    Keywords: affinity chromatography ; high-performance liquid chromatography ; bacteriophage T4 tail sheath protein ; bacteriophage T4 tail tube protein ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A novel method useful for selective isolation of the C-terminal peptide from a tryptic digestion mixture of a protein has been developed by taking advantage of a unique property of anhydrotrypsin, which has a strong specific affinity for the peptides containing arginine or lysine at their C-termini. Briefly, peptides produced by tryptic digestion of a protein are fractionated by affinity chromatography on a column of immobilized anhydrotrypsin. The C-terminal peptide is recovered in a breakthrough fraction, which the remainders are adsorbed on the column (unless the protein ends in arginine or lysine). The breakthrough fraction is then subjected to reversed-phase high-perfomance liquid chromatography in order to purify the C-terminal peptide. Using this method, we have successfully isolated the C-terminal peptides from tryptic digests of the sheath protein (gp 18) and the tube protein (gp 19) of bacteriophage T4. The analytical results on these peptides, together with the information on the N-terminal structures of the original proteins and on the nucleotide sequences of genes 18 and 19, allowed us to establish the complete primary structures of the two proteins.
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  • 81
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 109-115 
    ISSN: 0887-3585
    Keywords: proteins ; protein dynamics ; tryptophan exposure ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Experiments were done to test the thesis that acrylamide and similar small molecules can penetrate into proteins on a nanosecond time scale. The approach taken was to measure the pattern of fluorescence quenching exhibited by quenching molecules differing in molecular character (size, polarity, charge) when these are directed against protein tryptophans that cover the whole range of tryptophan accesibility. If quenching involves protein penetration and internal quencher migration, one expects that larger quenchers and more polar quenchers should display lesser quenching. In fact, no significant dependence on quencher character was found. For proteins that display measurable quenching, the disparate quenchers studied display very similar quenching rate constants when directed against any particular protein tryptophan. For several proteins having tryptophans known to be buried, no quenching occurs. These results are not consistent with the view that the kinds of small molecules studied can quite generally penetrate into and diffuse about within proteins at near-diffusion-limited rates. Rather the results suggest that when quenching is observed, the pathway involves encounters with tryptophans that are partially exposed at the protein surface. Available crystallographic results support this conclusion.
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  • 82
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 116-124 
    ISSN: 0887-3585
    Keywords: translational repressor ; in vitro transcription-translation ; gene expression ; protein purification ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The bacteriophage T4 translational represor regA protein has been purified from an overproducing strain, and its activity has been studied in simple in vitro protein synthesis reactions. RegA protein was found to inhibit the translation of T4 genes 44, 45, and ORF45-1 in a concentration-dependent fashion. Expression of two other T4 genes which are insensitive to regA protein in vivo, genes 32 and 43, was unaffected by the presence of regA protein. Specific inhibition of synthesis of genes 44, 45, and ORF 45-1 proteins was achieved with 5-20 μM concentrations of regA protein, without the addition of any other T4 encoded proteins or cofactors. When in vitro protein synthesis was performed in two steps, uncoupling translation from transcription, regA protein had an inhibitory effect regardless of whether it was added at the initiation of transcription or only at the translation step. This indicates that regA protein functions during the translation step of protein synthesis in vitro in agreement with previous in vivo studies of regA protein.
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  • 83
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 125-133 
    ISSN: 0887-3585
    Keywords: regulation ; prokaryotic bacteria ; leucine uptake ; leucyl tRNA corepressor ; cell physiology ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The livR gene encoding the repressor for high-affinity branched-chain amino acid transport in Escherichia coli has been cloned from a library prepared from the episome F106. The inserted DNA fragment from the initial cloned plasmid, pANT1, complemented two independent, spontaneously derived, regulatory mutations. Subcloning as well as the creation of deletions with Bal31 exonuclease revealed that the entire regulatory region is contained within a 1.1-kb RsaI-SalI fragment. Expression of the pANT plasmids in E. coli minicells showed that the regulatory region encodes one detectable protein with an apparent molecular weight of 21,000. DNA sequencing revealed one open reading frame of 501 bp encoding a protein with a calculated MW of 19,155. The potential secondary structure of the regulatory protein has been predicted and it suggests that the carboxy terminus may fold into three consecutive alpha helices. These results suggests that the livR gene encodes a repressor which plays a role in the regulation of expression of the livJ and the livK transport genes.
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  • 84
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 134-138 
    ISSN: 0887-3585
    Keywords: enzymatic transesterification ; peptide synthesis ; trypsin ; chymotrypsin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Trypsin and α-chymotrypsin were immobilized to alumina-phosphocolamine complex, activated by glutaraldehyde. The immobilized enzymes show a great stability toward organic solvents miscible or immiscible with water. In the presence of a low concentration of water, the immobilized enzymes catalyzed transesterification reactions as well as peptide synthesis. The synthesized peptides were stable toward the immobilized enzymes.
    Additional Material: 3 Ill.
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  • 85
    ISSN: 0887-3585
    Keywords: hypertension ; renin production ; mammalian expression ; affinity chromatography ; genetic engineering ; prorenin secretion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Transfection of Chinese hamster ovary (CHO) cells with a plasmid containing the cDNA for human preprorenin has provided cell lines that secrete 15 mg of native prorenin per liter of culture medium. Tryptic activation of the prorenin occurs by selective cleavage of the Arg66-Leu67 bond (numbering as in preprorenin). The renin product, purified in a single step and in high yield by affinity chromatography, is fully stable for as long as 8 months when stored in solution at 4°C and pH 6.5. Purity of the renin was judged to be greater than 95% by gel electrophoresis, compositional and N-terminal sequence analyses, and specific enzyme activity. An important aspect of the present work is the development of a direct assay for renin which permits accurate and reproducible evaluation of enzyme units and kinetic parameters. Application of methods described herein, combined with appropriate scale-up fermentation capabilities, provides the means for generating gram quantities of human renin and its zymogen.
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  • 86
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 146-163 
    ISSN: 0887-3585
    Keywords: protein conformation ; energy calculations ; protein modeling ; loop conformation ; surface area ; homologous proteins ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The feasibility of determining the conformation of segments of polypeptide chain up to six residues in length in globular proteins by means of systematic search through the possibles conformations has been investigated. Trial conformations are generated by using representative sets of φ, ψ, and χ angels that have been derived from an examination of the distributions of these angles in refined protein structures. A set of filters based on simple rules that protein structures obey is used to reduce the number of conformations to a manageable total. The most important filters are the maintenance of chain integrity and the avoidance of tooshort van der Waals contacts with the rest of the protein and with other portions of the segment under construction. The procedure is intended to be used with approximate models so that allowance is made throughout for errors in the rest of the structure. All possible main chains are first constructed and then all possible side-chain conformations are built onto each of these. The electrostatic energy, including a solvent screening term, and the exposed hyrophobic area are evaluated for each accepted conformation. The method has been tested on two segments of chain in the trypsin like enzyme from Streptomyces griseus. It is found that there is a wide spread of energies among the accepted conformations, and the lowest energy ones have satisfactorily small root mean square deviations from the X-ray structure.
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  • 87
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 164-175 
    ISSN: 0887-3585
    Keywords: DPG ; organophosphates ; ligand interactions ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Rate equilibrium dialysis was used to measure the binding of 2,3-diphosphoglycerate (DPG) to human oxy- and deoxyhemoglobin AO over the range pH 5-9, at 21.5°C. This approach yielded an accurate, precise, and self-consistent set of model-independent association constants. These data were successfully fitted to a thermodynamic model which is fuctionally similar to a Hill equation. The isotherms generated by this fitting procedure appear to intersect at low pH and converge at high pH. This apparent convergence at high pH is consistent with results obtained by oxygen equilibria studies performed under conditions of saturating DPG. These calculated isotherms were used to determine the enhancement of the Bohr effect as a function of pH. These results are consistent with data obtained by pH stat measurements by other investigators.This paper presents the first in a series of studies that will provide a systematic characterization of the interaction between hemoglobin and DPG.
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  • 88
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 176-187 
    ISSN: 0887-3585
    Keywords: drifted micrographs ; rotational blur ; thin sections ; Wiener filtering ; image analysis ; sickle hemoglobin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: We have investigated the restoration of electron micrographs exhibiting blurring due to drift and rotation. Blurring due to drift arises in micrographs taken of a specimen which is moving relative to the image plane. A related problem is that of rotational blurring which arises in micrographs of thin sections of helical particles viewed in cross section. The twist of the particle within the finite thickness of the section causes the image to appear rotationally blurred about the helical axis.Restoration algorithms were evaluated by applying them to the restoration of blurred model images degraded by additive Gaussian noise. Model images were also used to investigate how an incorrect estimate of the point spread function function describing the blur would effect the restoration. Images were, if necessary, geometrically transformed to a space in which the point spread function of the blur can be considered as linear and space invariant as, under these conditions, the restoration algorithms are greatly simplified. In the case of the rotationally blurred images this procedur was accomplished by transforming the image to polar coordinates.The restoration techniques were successfully applied to blurred micrographs of bacteriophage T4 and crystals of catalase. The quality of the restoration was judged by comparisons of the restored images to undegraded images. Application to micrographs of rotationally blurred cross sections of helical macrofibers of sickle hemoglobin resulted in a reduction in the amount of rotational blurring.
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  • 89
    ISSN: 0887-3585
    Keywords: G-protein ; phototransduction ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The GTP-binding subunit of transducin (Tα) activates the cGMP phosphodiesterase (PDE) of bovine retinal rods by relieving the constraint imposed by the inhibitory subunit PDEγ. We have isolated and characterized the complex Tα.GTPγS-PDEγ formed when Tα is activated by the nonhydrolyzable analog GTPγS. Sedimentation and light-scattering techniques demonstrate that, in contrast to free Tγ.GTPγS, which is soluble, the Tα.GTPγS-PDEγ complex, as well as Tα.GTP-PDEγ, is membrane bound at cytosolic ionic strength. It is eluted from the membrane at low ionic strength as a monomeric and 1:1 stoichiometric complex. The relative affinities of PDEγ for PDEαβ and for Tα.GTP are discussed.
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  • 90
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    Proteins: Structure, Function, and Genetics 1 (1986) 
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
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  • 91
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 195-210 
    ISSN: 0887-3585
    Keywords: protein domains ; oligomeric assembly ; RNA splicing ; multiple binding sites ; RNP core proteins ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Additional Material: 2 Ill.
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  • 92
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 211-217 
    ISSN: 0887-3585
    Keywords: protein folding ; α-helix stabilization ; peptide structural stability ; circular dichroism ; protein electrostatics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The effects of trifluoroethanol (TFE) on the stability of the α-helix formed by ribonuclease S-peptide, residues 1-19 of ribonuclease A, were studied by measuring circular dichroism as a function of TFE concentration, pH, and temperature. The S-peptide forms an unusually stable α-helix, which is known to be stabilized by TFE. The magnitude of the effect of charged groups on the peptide, manifested by the change in α-helix stability as a function of pH, was not altered significantly by either TFE concentration or temperature, indicating that the lower dielectric constant of TFE is not important in the stabilization of this α-helix. This suggests that the α-helix might be stabilized by many interactions in addition to the effects of charges. The titration curve of circular dichroism vs. TFE concentration appears to be cooperative at 0°C, but becomes progressively less cooperative at temperatures between 25 and 75°C. The properties of the TFE stabilization indicate that TFE might be a useful probe with which to measure the stability of marginally stable peptides and small proteins.
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  • 93
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 230-238 
    ISSN: 0887-3585
    Keywords: microcins ; peptide antibiotic ; protein processing ; HPLC ; protein secretion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Microcin B17 is a low-molecular-weight protein that inhibits DNA replication in a number of enteric bacteria. It is produced by bacterial strains which harbor a 70-kilobase plasmid called pMccB17. Four plasmid genes (named mcbABCD) are required for its production. The product of the mcbA gene was identified by labelling minicells. The mcbA gene product was slightly larger when a mutation in any of the other three production genes was present. This indicates that these genes are involved in processing the primary mcbA product to yield the active molecule. The mcbA gene product predicted from the nucleotide sequence has 69 amino acids including 28 glycine residues. Microcin B17 was extracted from the cells by boiling in 100 mM acetic acid, 1 mM EDTA, and purified to homogeneity in a single step by high-performance liquid chromatography through a C18 column. The N-terminal amino acid sequence and amino acid composition demonstrated that mcbA is the structural gene for microcin B17. The active molecule is a processed product lacking the first 26 N-terminal residues. The 43 remaining residues include 26 glycines. While microcin B17 is an exported protein, the cleaved N-terminal peptide does not have the characteristic properties of a “signal sequence,” which suggests that it is secreted by a mechanism different from that used by most secreted proteins of E. coli.
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  • 94
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 239-246 
    ISSN: 0887-3585
    Keywords: halobacteria ; photosensory receptor ; retinal ; slow-cycling rhodopsins ; sensory rhodopsins ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A second slow-cycling retinylidene protein, in addition to slow-cycling (sensory) rhodopsin (SR), can be bleached with hydroxylamine and regenerated with all-trans retinal in photosensory signaling Halobacterium halobium membranes. Flash photolysis shows this protein undergoes a photochemical reaction cycle characterized by photoconversion of its ground state (λmax 480 nm) to a species with λmax ≤ 360 nm, which thermally regenerates the 480-nm species with a t½ of 260 msec at 25°C, under conditions in which SR photocycles at 650 msec in the same membranes. Mutants characterized with respect to their phototaxis behavior are identified which contain SR and the 480-nm pigment, the latter ranging from undetectable to a concentration equal to that of SR. Receptor mutants lacking all phototaxis sensitivity lack both of the photochemically reactive proteins. The mutant properties contribute to an accumulation of behavioral and spectroscopic evidence that the 480-nm pigment is a second sensory photoreceptor in H. halobium. NaDodSO4-polyacrylamide gel electrophoresis of [3H]retinal-labeled membrane proteins from the mutants indicates SR and the 480-nm pigment contain distinct chromophoric polypeptides differing in their migration rates. The data implicate polypeptides of 25,000 Mr and 23,000 Mr as retinal-binding polypeptides of SR and the 480-nm protein, respectively.
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  • 95
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 218-229 
    ISSN: 0887-3585
    Keywords: colicin E1 ; site-directed mutagenesis ; ion channel ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Cleavage of colicin E1 molecules with a variety of proteases or with cyanogen bromide (CNBr) generates COOH-terminal fragments which have channel-forming activity similar to that of intact colicin in planar lipid bilayer membranes. The smallest channel-forming fragment obtained by CNBr cleavage of the wild-type molecule consists of the C-terminal 152 amino acids. By the use of oligonucleotide-directed mutagenesis, we have made nine mutants along this 152 amino acid peptide, in which an amino acid was replaced by methionine in order to create a new CNBr cleavage site. The smallest of the CNBr-cleaved C-terminal fragments with channel-forming activity, in planar bilayer membranes, was generated by cleavage at new Met position 428 and has 94 amino acids, whereas a 75 amino acid peptide produced by cleavage of a new Met at position 447 did not have channel activity. The NH2-terminus of the channel-forming domain of colicin E1 appears therfore to lie between residues 428 and 447. Since, however, the last six C-terminal residues of the colicin can be removed without changing activity, the number of amino acids necessary to form the channel is 88 or less. In addition, the unique Cys residue in colicin E1 was replaced by Gly, and nine mutants were then made with Cys placed at sequential locations along the peptide for eventual use as sulfhydryl attachment sites to determine the local environment of the replaced amino acid. In the course of making 21 mutants, eight charged residues have been replaced by uncharged Met or Cys without changing the biological activity of the intact molecule.It has been proposed previously that the conformation of the colicin E1 channel is a barrel formed from five or six α-helices, each having 20 amino acids spanning the membrane and two to four residues making the turn at the boundary of the membrane. Our finding that 88 amino acids can make an active channel, combined with recently reported stoichiometric evidence that the channel is a monomer excludes this model and adds significant constraints which can be used in building a molecular model of the channel.
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  • 96
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 247-255 
    ISSN: 0887-3585
    Keywords: protein folding ; domain interactions ; fluorescence transfer ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: This report describes the use of fluorescence energy transfer between an intrinsic energy donor (tryptophan 177) and two chemically added acceptors to study intermediates in the folding of the β2 subunit of E. coli tryptophan-synthase. Two early folding steps are thus identified and characterized. One is very rapid (its rate constant at 12°C is 0.02 sec-1) and corresponds to the folding of the N-terminal domain into a structure whose overall features approximate well those of the native domain. The second step is somewhat slower (its rate constant at 12°C is 0.008 sec-1) and involves a conformational rearrangement of the N-terminal domain brought about by the interactions between the N-and C-terminal domains within a monomeric β chain. This brings to five the number of intermediates which have been identified and ordered on the folding pathway of the dimeric β2 subunit.
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  • 97
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 263-266 
    ISSN: 0887-3585
    Keywords: protein-DNA interaction ; overproducer clone ; sequence-specific recognition ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: HhaII restriction endonuclease purified from an overproducing recombinant E. coli clone has been cocrystallized with a heptanucleotide duplex, d-GGAGTCC:GGACTCC. The cocrystals are monoclonic and belong to the space group C2. The unit cell dimensions are a = 199.0±1.0 Å, b = 100.0±0.5 Å, c = 80.3±0.4 Å, and β = 101.0±1.0°. There appear to be two dimers per asymmetric unit and the crystals diffract to 4-Å resolution.
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  • 98
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    Proteins: Structure, Function, and Genetics 1 (1986), S. 267-279 
    ISSN: 0887-3585
    Keywords: molecular model building ; energy minimization ; homology modeling ; site-specific mutagenesis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A procedure (coupled perturbation procedure, CPP) is introduced as a specific method for calculating the detailed three-dimensional structure of a protein molecule which has a nummber of amino-acid substitutions relative to some previously determined “parent” protein structure. The accuracy of the procedure is tested by calculating the conformation of a region of the human immunoglobulin fragment Fab Kol based on the analogous region of the human immunoglobulin fragment Fab New. Both structures have previously been determined crystallographically. The calculated model is accurate to the extent that both of the sequence differences in the region are modeled correctly and that conformational changes in a number of nearby residues are correctly identified. CPP is shown to give better results than other commonly used modeling procedures when applied to the same problem.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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  • 99
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 256-262 
    ISSN: 0887-3585
    Keywords: FV fragment ; space filling hapten ; reconstitution ; scratched analysis ; equilibrium dialysis ; contact residues ; hypervariable loops ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: IgG Gar, a human myeloma protein that binds riboflavin with a high affinity, was used to derive variable region fragments from the heavy chain and the light chain. Riboflavin binding ability of the active site generated by V(H) and light chain and the active site generated by V(H) and V(L) was compared to riboflavin binding by the F(ab) fragment. The riboflavin binding ability of the F(ab) fragment is the same as the intact molecule, while the binding ability of the active site formed by V(H) and light chain is lowered by two to three orders of magnitude, indicating that the removal of C(H1) domain decreases the interaction between riboflavin and the amino acids that is important in tight binding of riboflavin. Removal of the third hypervariable region and the constant region domain from the light chain further lowers the binding constant by one order of magnitude. The results indicate that the V(H) and V(L) segments of IgG Gar can reconstitute a riboflavin binding site. The decrease in affinity probably reflects a decrease in the rigidity with which the hypervariable loops are held together to place the contact amino acid residues in optimal contact with the hapten.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 100
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 280-286 
    ISSN: 0887-3585
    Keywords: antibody affinity ; ELISA ; β-endorphin models ; melittin model ; amphiphilic α-helix ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Monoclonal antibodies against human apolipoprotein A-I (apoA-I) were generated by the hybridoma technique. Clone G-10 was selected on the basis of its highest titer. The affinity of this antibody toward a series of synthetic peptides differing in length, amino acid composition, and amphiphilicity was tested by using both the indirect and the competitive enzyme-linked immunosorbent techniques (ELISA). From these measurements we calculated dissociation constants of the complexes of the antibody with apoA-I bound to the surface of the microtiter plate, apoA-I in solution, and any of the several peptides in solution. The dissociation constant (Kd) of the immobilized apoA-I/anti-apoA-I-complex, Kd = 2 × 10-9 M, was significantly lower than that of the complex resulting from the interaction between anti-apoA-I and either apoA-I in solution or any of the several amphiphilic helical peptides in solution. Peptides devoid of amphiphilic secondary structure were inert. These data are consistent with the proposal that monoclonal G-10 recognizes in antigenic peptides an α-helical secondary structure of defined hydrophilic-lipophilic balance and comparatively less the specific amino acid side chains. We propose that the highest contribution to the free energy of binding (8 Kcal/mole) is derived from the docking of the helix to the antibody. It follows that in probing the specificity of a monoclonal antibody the conformation and the physical environment of the interacting antigen must be taken into account.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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